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CHMI 2227E Biochemistry I

CHMI 2227E Biochemistry I. Nucleic acids: structure physico-chemical properties. Nucleic acids. 32 P in nucleic acids only. 35 S in proteins only (Met/Cys). Hershey and Chase The Waring blender experiment…. Nature of nucleic acids 1. nitrogenated bases. Nature of nucleic acids 2. sugars.

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CHMI 2227E Biochemistry I

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  1. CHMI 2227EBiochemistry I Nucleic acids: structure physico-chemical properties CHMI 2227 - E.R. Gauthier, Ph.D.

  2. Nucleic acids CHMI 2227 - E.R. Gauthier, Ph.D.

  3. 32P in nucleic acids only 35S in proteins only (Met/Cys) Hershey and ChaseThe Waring blender experiment… CHMI 2227 - E.R. Gauthier, Ph.D.

  4. Nature of nucleic acids1. nitrogenated bases CHMI 2227 - E.R. Gauthier, Ph.D.

  5. Nature of nucleic acids2. sugars CHMI 2227 - E.R. Gauthier, Ph.D.

  6. Glycosidic bond H 9 1 H 1’ 1’ Deoxyadenosine Cytidine Nature of nucleic acids3. nucleosides CHMI 2227 - E.R. Gauthier, Ph.D.

  7. Nature of nucleic acids3. nucleosides CHMI 2227 - E.R. Gauthier, Ph.D.

  8. Nature of nucleic acids3. nucleosides CHMI 2227 - E.R. Gauthier, Ph.D.

  9. Beta b Gamma g Alpha a Deoxythymidine 5’triphosphate Guanosine 5’monophosphate Nature of nucleic acids4. nucleotides CHMI 2227 - E.R. Gauthier, Ph.D.

  10. Nature of nucleic acids4. nucleotides CHMI 2227 - E.R. Gauthier, Ph.D.

  11. Nature of nucleic acids5. polynucleotides CHMI 2227 - E.R. Gauthier, Ph.D.

  12. g b a http://www.mbi.ufl.edu/facilities/msg/bch4024m/lecture45.pdf Nature of nucleic acids5. phosphodiester bond CHMI 2227 - E.R. Gauthier, Ph.D.

  13. Nature of nucleic acids6. Chargaff’s rules CHMI 2227 - E.R. Gauthier, Ph.D.

  14. bases Sugar/phosphate backbone Structure of nucleic acids1. DNA CHMI 2227 - E.R. Gauthier, Ph.D.

  15. Structure of nucleic acids1. DNA CHMI 2227 - E.R. Gauthier, Ph.D. 1 Å (Ångstrom) = 0.1 nm = 1 x 10-10 m

  16. Structure of nucleic acids1. DNA CHMI 2227 - E.R. Gauthier, Ph.D.

  17. Structure of nucleic acids2. DNA/RNA hybrids and RNA duplexes CHMI 2227 - E.R. Gauthier, Ph.D.

  18. Structure of nucleic acids2. DNA/RNA hybrids and RNA duplexes CHMI 2227 - E.R. Gauthier, Ph.D.

  19. Structure of nucleic acids • Java applets: • http://www.moleculesinmotion.com/ CHMI 2227 - E.R. Gauthier, Ph.D.

  20. DNA absorbance CHMI 2227 - E.R. Gauthier, Ph.D.

  21. DNA denaturation CHMI 2227 - E.R. Gauthier, Ph.D.

  22. DNA#1 Hyperchromic shift DNA#2 DNA denaturation CHMI 2227 - E.R. Gauthier, Ph.D.

  23. G+C-rich A+T-rich DNA denaturation CHMI 2227 - E.R. Gauthier, Ph.D.

  24. Hybridization CHMI 2227 - E.R. Gauthier, Ph.D.

  25. - + Power • Polymerized agarose is porous, allowing for the movement of DNA Scanning Electron Micrograph of Agarose Gel (1×1 µm)  Agarose gel electrophoresis DNA CHMI 2227 - E.R. Gauthier, Ph.D.

  26. Staining with ethidium bromide Standard curve: Log length vs distance migrated Ethidium bromide fluoresces red only when it intercalates between base pairs. DNA molecules of known length Agarose gel electrophoresis CHMI 2227 - E.R. Gauthier, Ph.D.

  27. Annealing 32P-labeled nucleic acid probe is complementary to one of the DNA molecules on the gel Southern blot and Northern blot CHMI 2227 - E.R. Gauthier, Ph.D.

  28. Genomic DNA cut with restriction enzyme Different allele? Southern blot CHMI 2227 - E.R. Gauthier, Ph.D.

  29. FISHFluorescent in-situ hybridization CHMI 2227 - E.R. Gauthier, Ph.D.

  30. RNA labeling 1000’s of genes Microarrays aka DNA chips CHMI 2227 - E.R. Gauthier, Ph.D.

  31. DNA denaturation CHMI 2227 - E.R. Gauthier, Ph.D.

  32. DNA denaturation CHMI 2227 - E.R. Gauthier, Ph.D.

  33. The «C value» paradox CHMI 2227 - E.R. Gauthier, Ph.D.

  34. The fruit fly Drosophila melanogaster The mountain grasshopper Podisma pedestris 180 Mb 18,000 Mb kbp The «C value» paradox CHMI 2227 - E.R. Gauthier, Ph.D.

  35. Highly repeated Moderately repeated Single copy The «C value» paradox CHMI 2227 - E.R. Gauthier, Ph.D.

  36. Human genome Human genome 3000 Mb Genes and other related sequences 900 Mb Extra-genic DNA 2100 Mb Coding DNA 90 Mb Non-coding DNA 810 Mb Repeated DNA 420 Mb Single/low copy number DNA 1680 Mb Tandem repeats Interspersed repeats Pseudogenes Gene fragments Introns, etc Microsatellite DNA Satellite DNA LTR elements SINE elements CHMI 2227 - E.R. Gauthier, Ph.D. Minisatellite DNA LINE elements Transposons

  37. Human genome CHMI 2227 - E.R. Gauthier, Ph.D.

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